Crystallization and preliminary X-ray diffraction analysis of the HsdR subunit of a putative type I restriction enzyme from Vibrio vulnificus YJ016.
Type I restriction enzymes are multimeric proteins that consist of three subunits. The HsdS and HsdM subunits form a complex protein that shows methyltransferase activity, while the HsdR subunit functions as an endonuclease as well as as a translocase. Of these three subunits, no structural information on the HsdR subunit is yet available. The putative HsdR gene from Vibrio vulnificus YJ016 (HsdR_Vv) was cloned and expressed and the expressed protein HsdR_Vv was purified. HsdR_Vv was crystallized from 8%(w/v) polyethylene glycol 3350, 0.15 M ammonium chloride, 0.1 M HEPES pH 7.5 and 2 mM beta-mercaptoethanol. Diffraction data were collected to 2.60 A resolution using synchrotron radiation. The crystal belongs to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 71.01, b = 89.04, c = 113.66 A. With one HsdR_Vv molecule in the asymmetric unit, the Matthews coefficient was 2.14 A(3) Da(-1) and the solvent content was 42%.
Uyen
Nguyen To
NT
Interdisciplinary Graduate Program in Molecular Medicine, Gwangju 501-746, Republic of Korea.
Nishi
Kosuke
K
Park
Suk Youl
SY
Choi
Ji Woo
JW
Lee
Hyun Ju
HJ
Kim
Jeong Sun
JS
D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D003460:Crystallization; D004262:DNA Restriction Enzymes / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D015253:Deoxyribonucleases, Type I Site-Specific / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D021122:Protein Subunits / Q000737:chemistry* / Q000378:metabolism*; D041261:Vibrio vulnificus / Q000201:enzymology*; D014961:X-Ray Diffraction
D001426:Bacterial Proteins; D021122:Protein Subunits; D004262:DNA Restriction Enzymes; D015253:Deoxyribonucleases, Type I Site-Specific